Beijing Institute of Basic Medical Sciences, Beijing 100850, China; State Key Laboratory of Natural Medicines, School of Life Science and Technology, China Pharmaceutical University, Nanjing, Jiangsu 211198, China; Research Center for Biochemistry and Molecular Biology, Xuzhou Medical University, Xuzhou, Jiangsu 221004, China.
Beijing Institute of Basic Medical Sciences, Beijing 100850, China.
Biomed Pharmacother. 2024 Apr;173:116342. doi: 10.1016/j.biopha.2024.116342. Epub 2024 Mar 1.
Neuroinflammation is responsible for neuropsychiatric dysfunction following acute brain injury and neurodegenerative diseases. This study describes how a hypoxia-inducible factor prolyl hydroxylase (HIF-PHD) inhibitor FG-4592 prevents the lipopolysaccharide (LPS)-induced acute neuroinflammation in microglia.
The distribution of FG-4592 in mouse brain tissues was determined by collision-induced dissociation tandem mass spectrometry. Microglial activation in the hippocampus was analyzed by immunofluorescence. Moreover, we determined the activation of HIF-1 and nuclear factor-κB (NF-κB) signaling pathways, proinflammatory responses using molecular biological techniques. Transcriptome sequencing and BNIP3 silencing were conducted to explore signaling pathway and molecular mechanisms underlying FG-4592 anti-inflammatory activity.
FG-4592 was transported into the brain tissues and LPS increased its transportation. FG-4592 promoted the expression of HIF-1α and induced the downstream gene transcription in the hippocampus. Administration with FG-4592 significantly inhibited microglial hyperactivation and decreased proinflammatory cytokine levels following LPS treatment in the hippocampus. The LPS-induced inflammatory responses and the NF-κB signaling pathway were also downregulated by FG-4592 pretreatment in microglial cells. Mechanistically, Venn diagram analysis of transcriptomic changes of BV2 cells identified that BNIP3 was a shared and common differentially expressed gene among different treatment groups. FG-4592 markedly upregulated the protein levels of BNIP3 in microglia. Importantly, BNIP3 knockdown aggravated the LPS-stimulated inflammatory responses and partially reversed the protection of FG-4592 against microglial inflammatory signaling and microglial activation in the mouse hippocampus.
FG-4592 alleviates neuroinflammation through facilitating microglial HIF-1/BNIP3 signaling pathway in mice. Targeting HIF-PHD/HIF-1/BNIP3 axis is a promising strategy for the development of anti-neuroinflammation drugs.
神经炎症是导致急性脑损伤和神经退行性疾病后神经精神功能障碍的原因。本研究描述了缺氧诱导因子脯氨酰羟化酶（HIF-PHD）抑制剂 FG-4592 如何防止脂多糖（LPS）诱导小胶质细胞中的急性神经炎症。
通过碰撞诱导解离串联质谱法测定 FG-4592 在小鼠脑组织中的分布。通过免疫荧光分析海马体中的小胶质细胞活化。此外，我们使用分子生物学技术确定 HIF-1 和核因子-κB（NF-κB）信号通路、促炎反应的激活情况。进行转录组测序和 BNIP3 沉默，以探索 FG-4592 抗炎活性的信号通路和分子机制。
FG-4592 被转运到脑组织中，LPS 增加了其转运。FG-4592 促进了海马体中 HIF-1α 的表达，并诱导了下游基因的转录。FG-4592 预处理可显著抑制 LPS 处理后海马体中小胶质细胞的过度激活和促炎细胞因子水平的降低。FG-4592 预处理还可下调 LPS 诱导的小胶质细胞炎症反应和 NF-κB 信号通路。在机制上，BV2 细胞转录组变化的 Venn 图分析表明，BNIP3 是不同处理组之间共同的差异表达基因。FG-4592 显著上调了小胶质细胞中 BNIP3 的蛋白水平。重要的是，BNIP3 敲低加重了 LPS 刺激的炎症反应，并部分逆转了 FG-4592 对小胶质细胞炎症信号和小鼠海马体中小胶质细胞激活的保护作用。
FG-4592 通过促进小鼠小胶质细胞中的 HIF-1/BNIP3 信号通路来减轻神经炎症。靶向 HIF-PHD/HIF-1/BNIP3 轴是开发抗神经炎症药物的有前途的策略。
